Inhibitory effects of French pine bark extract, Pycnogenol®, on alveolar bone resorption and on the osteoclast differentiation.

Pycnogenol(®) (PYC) is a standardized bark extract from French maritime pine (Pinus pinaster Aiton). We examined the inhibitory effects of PYC on alveolar bone resorption, which is a characteristic feature of periodontitis, induced by Porphyromonas gingivalis (P. gingivalis) and osteoclast differentiation. In rat periodontitis model, rats were divided into four groups: group A served as the non-infected control, group B was infected orally with P. gingivalis ATCC 33277, group C was administered PYC in the diet (0.025%: w/w), and group D was infected with P. gingivalis and administered PYC. Administration of PYC along with P. gingivalis infection significantly reduced alveolar bone resorption. Treatment of P. gingivalis with 1 µg/ml PYC reduced the number of viable bacterial cells. Addition of PYC to epithelial cells inhibited adhesion and invasion by P. gingivalis. The effect of PYC on osteoclast formation was confirmed by tartrate-resistant acid phosphatase staining. PYC treatment significantly inhibited osteoclast formation. Addition of PYC (1-100 µg/ml) to purified osteoclasts culture induced cell apoptosis. These results suggest that PYC may prevent alveolar bone resorption through its antibacterial activity against P. gingivalis and by suppressing osteoclastogenesis. Therefore, PYC may be useful as a therapeutic and preventative agent for bone diseases such as periodontitis.

Direct assessment of the antioxidant properties of midazolam by electron spin resonance spectroscopy.

Some antioxidant anesthetics directly inhibit lipid peroxidation mediated via the generation of reactive oxygen species (ROS). To date, the scavenging effects of midazolam on ROS have not been directly assessed. We investigated the inhibitory effect of midazolam on ROS [hydroxyl radical (HO(·)) and superoxide (O (2) (·-) )] by in vitro X-band electron spin resonance with the spin-trapping agent 5,5-dimethyl-1-pyrroline-N-oxide. Our results indicated that HO(·) and O (2) (·-) were not affected by midazolam at clinically relevant concentrations, but were directly scavenged by midazolam at high concentrations (i.e., >4.6 and >1.5 mM, respectively).

Microcirculation changes in gingival tissue after ultrasonic tooth preparation in beagle dogs.

OBJECTIVE: Ultrasonic wave technology is widely used during dental treatments. We previously demonstrated that this method protects the gingival tissue. However, the physiological change on the gingival microvasculature caused by this method remains unclear. The aim of this study was to investigate the relationship between the morphological and physiological effects on gingival microcirculation when preparing teeth, using the conventional dental turbine or ultrasonic method. METHODOLOGY: The lower premolar teeth of beagle dogs were prepared along the gingival margin by using a dental turbine or ultrasonic wave instrument. Gingival vasculature changes were investigated using scanning electron microscopy for corrosion resin casts. Gingival blood flow at the preparation site was determined simultaneously by laser Doppler flowmetry. These assessments were performed immediately (Day 0), at 7 days and 30 days after tooth preparation. RESULTS: At day 0, in the turbine group, blood vessels were destroyed and some resin leaked. Furthermore, gingival blood flow at the site was significantly increased. In contrast, the ultrasonic group demonstrated nearly normal vasculature and gingival blood flow similar to the non-prepared group for 30 days after preparation. No significant alterations occurred in gingival circulation 30 days after either preparation; however, the turbine group revealed obvious morphological changes. CONCLUSIONS: Based on multiple approach analyses, this study demonstrated that ultrasonic waves are useful for microvascular protection in tooth preparation. Compared with a dental turbine, ultrasonic wave instruments caused minimal damage to gingival microcirculation. Tooth preparation using ultrasonic wave instruments could be valuable for protecting periodontal tissue.

A Study of Deposition Coatings Formed by Electroformed Metallic Materials.

Major joining methods of dental casting metal include brazing and laser welding. However, brazing cannot be applied for electroformed metals since heat treatment could affect the fit, and, therefore, laser welding is used for such metals. New methods of joining metals that do not impair the characteristics of electroformed metals should be developed. When new coating is performed on the surface of the base metal, surface treatment is usually performed before re-coating. The effect of surface treatment is clinically evaluated by peeling and flex tests. However, these testing methods are not ideal for deposition coating strength measurement of electroformed metals. There have been no studies on the deposition coating strength and methods to test electroformed metals. We developed a new deposition coating strength test for electroformed metals. The influence of the negative electrolytic method, which is one of the electrochemical surface treatments, on the strength of the deposition coating of electroformed metals was investigated, and the following conclusions were drawn: 1. This process makes it possible to remove residual deposits on the electrodeposited metal surface layer. 2. Cathode electrolysis is a simple and safe method that is capable of improving the surface treatment by adjustments to the current supply method and current intensity. 3. Electrochemical treatment can improve the deposition coating strength compared to the physical or chemical treatment methods. 4. Electro-deposition coating is an innovative technique for the deposition coating of electroformed metal.

Microcirculation changes in gingival tissue after ultrasonic tooth preparation in beagle dogs

Abstract Ultrasonic wave technology is widely used during dental treatments. We previously demonstrated that this method protects the gingival tissue. However, the physiological change on the gingival microvasculature caused by this method remains unclear. Objective The aim of this study was to investigate the relationship between the morphological and physiological effects on gingival microcirculation when preparing teeth, using the conventional dental turbine or ultrasonic method. Methodology The lower premolar teeth of beagle dogs were prepared along the gingival margin by using a dental turbine or ultrasonic wave instrument. Gingival vasculature changes were investigated using scanning electron microscopy for corrosion resin casts. Gingival blood flow at the preparation site was determined simultaneously by laser Doppler flowmetry. These assessments were performed immediately (Day 0), at 7 days and 30 days after tooth preparation. Results At day 0, in the turbine group, blood vessels were destroyed and some resin leaked. Furthermore, gingival blood flow at the site was significantly increased. In contrast, the ultrasonic group demonstrated nearly normal vasculature and gingival blood flow similar to the non-prepared group for 30 days after preparation. No significant alterations occurred in gingival circulation 30 days after either preparation; however, the turbine group revealed obvious morphological changes. Conclusions Based on multiple approach analyses, this study demonstrated that ultrasonic waves are useful for microvascular protection in tooth preparation. Compared with a dental turbine, ultrasonic wave instruments caused minimal damage to gingival microcirculation. Tooth preparation using ultrasonic wave instruments could be valuable for protecting periodontal tissue.

Porphyromonas gingivalis-induced alveolar bone loss is accelerated in the stroke-prone spontaneously hypertensive rat.

Porphyromonas gingivalis (P. gingivalis) is one of the prominent periodontal pathogens and is the most important bacteria involved in the onset and exacerbation of periodontitis. P. gingivalis is an anaerobic, Gram-negative coccobacillus that plays a role in the progression of periodontal disease by promoting alveolar bone resorption. The aim of the present study was to examine P. gingivalis-induced osteoclastic bone resorption in the stroke-prone spontaneously hypertensive rat (SHRSP), in which oxidative stress induced by reactive oxygen species (ROS) is increased. In the present study, we used animals orally challenged with P. gingivalis as a chronic inflammation model. Horizontal bone loss around the maxillary molars was assessed morphometrically. Animals were divided into four groups: (1) P. gingivalis-non-infected Wister Kyoto Rat (WKY), (2) orally challenged with P. gingivalis WKY (WKY + Pg), (3) P. gingivalis-non-infected SHRSP, and (4) orally challenged with P. gingivalis SHRSP (SHRSP + Pg). Alveolar bone resorption was significantly increased in the orally challenged with P. gingivalis groups, and was accelerated in the SHRSP group. Histological analysis revealed that the infiltration of inflammatory cells was absent in all groups. However, the infiltration of osteoclasts was observed in the SHRSP + Pg and SHRSP groups. We examined P. gingivalis-induced alveolar bone loss in both the SHRSP and WKY. The results obtained demonstrated that P. gingivalis-induced alveolar bone loss would be involved in hypertension and stroke animal model, such as SHRSP and/or periodontal disease.

Reactive oxygen species scavenging activity of Jixueteng evaluated by electron spin resonance (ESR) and photon emission.

Jixueteng, the dried stem of Spatholobus suberectus Dunn (Leguminosae), is a traditional Chinese herbal medicine that is commonly classified as a herb that promotes blood circulation and can be used to treat blood stasis. The aim of this study was to examine the reactive oxygen species (ROS) scavenging activity of Jixueteng and other herbal medicines. The ROS scavenging activities of the water extracts of Jixueteng, Cnidium officinale and Salvia miltiorrhiza were examined using an electron spin resonance (ESR) technique and faint luminescence measurement. The ESR signal intensities of the superoxide anion (O2·) and hydroxyl radical (HO·) were reduced more by Jixueteng than the other herbal medicines we tested. High photon emission intensity to hydrogen peroxide (H202) and HO· was observed in Jixueteng using the XYZ chemiluminescence system that was used as faint luminescence measurement and analysis. The results of the present study revealed that the ROS scavenging activity of 8% Jixueteng was the strongest among the herbal medicines we tested. It has been reported that Jixueteng includes various polyphenols. In the ROS scavenging activity by Jixueteng, it is supposed that the antioxidant activity caused by these polyphenols would contribute greatly. In conclusion, a water extract component of Jixueteng had potent free radical scavenging activity and an antioxidative effect that inhibited the oxidative actions of O2·â», H2O2 and HO·. Therefore, Jixueteng represents a promising therapeutic drug for reactive oxygen-associated pathologies.

Inhibitory effects of Jixueteng on P. gingivalis-induced bone loss and osteoclast differentiation.

OBJECTIVE: The aim of this study was to investigate the possibility of Jixueteng as a preventive and therapeutic drug for the periodontitis. We investigated the inhibitory effects of Jixueteng on Porphyromonas gingivalis-induced bone loss in mice, antibacterial activity against P. gingivalis and differentiation of osteoclast and viability of cells. MATERIALS AND METHODS: Fifty-four male, 4-week-old C57BL/6N mice, were randomly divided into the following three groups of 18 mice each; group A served as the P. gingivalis non-infected control (sham group), group B was infected orally with P. gingivalis and group C was administered Jixueteng extract in drinking water and was then infected with P. gingivalis. In order to evaluate the effect of Jixueteng, the distance from the alveolar bone crest to the cemento-enamel junction was determined. P. gingivalis suspension was exposed for 1, 15 and 60 min to 5 ml of the Jixueteng extract. Furthermore, to clarify the mechanism of the inhibitory effects of Jixueteng on osteoclast formation, Jixueteng extract was added to the culture of mouse bone marrow cells, osteoclast precursor. RESULTS: Administration of Jixueteng along with P. gingivalis infection significantly reduced alveolar bone loss compared to P. gingivalis infection. Jixueteng treatment at the concentration of 0.01% significantly inhibited osteoclast formation. The addition of Jixueteng extract (0.1%, 0.01%, and 0.001%) to the culture showed a significant inhibition of the number of surviving osteoclasts in a dose-dependent manner. CONCLUSION: Jixueteng has an antibacterial activity against P. gingivalis and inhibitory effects on osteoclastogenesis, it may be useful as a therapeutic drug in the treatment of P. gingivalis-induced periodontitis.

Gingival vascular functions are altered in type 2 diabetes mellitus model and/or periodontitis model.

The association of vascular reactivity between diabetes and periodontal disease has not been clarified. Gingival blood flow was measured by laser Doppler flowmetry for 31 weeks in Wistar rats, Wistar rats orally challenged with Porphyromonas gingivalis (Wistar rats + Porphyromonas gingivalis), Goto-Kakizaki rats, and Goto-Kakizaki rats orally challenged with Porphyromonas gingivalis (Goto-Kakizaki rats + Porphyromonas gingivalis). Effects of alveolar bone resorption on periodontal tissue was enhanced in Wistar rats + Porphyromonas gingivalis, and Goto-Kakizaki rats, with this effect being significantly enhanced by Goto-Kakizaki rats + Porphyromonas gingivalis. Using the L-band electron spin resonance technique, we succeeded in measuring oxidative stress as decay rate constant (K(1) and K(2)) of 3-carbamoyl-2,2,5,5-tetramethylpyrrolidin-1-yloxy in the oral and maxillofacial region of the animal models. The decay rate constant (K(1)) of 3-carbamoyl-2,2,5,5-tetramethylpyrrolidin-1-yloxy was significantly greater in the oral and maxillofacial region of Goto-Kakizaki rats + Porphyromonas gingivalis compared to Wistar rats, Wistar rats + Porphyromonas gingivalis and Goto-Kakizaki rats groups. Gingival reactive hyperemia was attenuated by periodontal disease, and this effect was also remarkable in the diabetes mellitus model. Taken together, we found that vascular endothelial function was decreased in diabetes mellitus and/or periodontal disease animal models due to increasing oxidative stress in the gingival circulation.

Assessments of salivary antioxidant activity using electron spin resonance spectroscopy.

OBJECTIVE: In recent years, the function of saliva has been focused on evaluation of general status. The relationship between salivary antioxidant activity and periodontal disease progression is unclear. The aim of this study is to assess the relationship between periodontal disease and salivary antioxidant activity towards various reactive oxygen species (ROS) using electron spin resonance (ESR) technique. METHODS: We demonstrated that whole saliva derived rats or human subjects scavenged ROS such as superoxide (O(2)(·-)) and hydroxyl radical (HO(·)) using ESR spectroscopy with spin trapping agent. In addition, we assessed the relationship between antioxidants activity towards ROS and periodontal index with superoxide dismutase (SOD) activity in human subject saliva. RESULTS: Antioxidant activity towards O(2)(·-) was increased by Porphyromonas gingivalis (P. gingivalis) infection in rat, although antioxidant activity towards HO(·) was not changed. In human, a strong correlation (r = 0.88, p < 0.01) recognized between salivary antioxidant activity towards O(2)(·-) and probing pocket depth (PPD). In addition, the intensity of salivary antioxidant activity depended on SOD activity level. SOD activity was also correlated with PPD. CONCLUSIONS: Rat salivary antioxidant activity towards O(2)(·-) was up-regulated by the inflammatory response caused by P. gingivalis infection. Similar response was recognized in human saliva with periodontal index. Additionally, a linear correlation between antioxidant activity towards O(2)(·-) and SOD activity was verified by ESR technique. Therefore, evaluation of the salivary antioxidant activity towards O(2)(·-) might be an effective parameter for the objective assessment of periodontal disease progression.